Pharmaceutical formulation

ABSTRACT

A pharmaceutical composition suitable for oral administration comprising particles of 2-(1,5-dimethyl-3-phenyl-1H-pyrrol-2-yl)-N-(4-(4-(5-fluoropyrimidin-2-yl)piperazin-1-yl)phenyl)-2-oxoacetamide is provided. Also provided is a pharmaceutical composition suitable for parenteral administration wherein the composition comprises 2-(1,5-dimethyl-3-phenyl-1H-pyrrol-2-yl)-N-(4-(4-(5-fluoropyrimidin-2-yl)piperazin-1-yl)phenyl)-2-oxoacetamide. The compositions are useful in the treatment of fungal infection in a subject in need thereof.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a continuation of U.S. patent application Ser. No.16/303,999, filed on May 25, 2017, which is a U.S. national phaseapplication under 35 U.S.C. § 371 of International Application No.PCT/GB2017/051494, filed on May 25, 2017, which claims priority benefitof British Patent Application No. 1609222.3, filed on May 25, 2016, thedisclosures of each of which are hereby incorporated by reference intheir entireties.

FIELD OF THE INVENTION

This invention relates to pharmaceutical formulations comprising apyrrole compound. Specifically, it relates to pharmaceuticalcompositions which are suitable for oral administration, topharmaceutical compositions which are suitable for parenteraladministration, and to the use of such compositions in prevention ortreatment of fungal diseases. It also relates to a method of producing apharmaceutical composition as described herein.

BACKGROUND OF THE INVENTION

Invasive fungal infections are well recognised as diseases of theimmunocompromised host. Over the last twenty years there have beensignificant rises in the number of recorded instances of fungalinfection. In part this is due to increased awareness and improveddiagnosis of fungal infection. However, the primary cause of thisincreased incidence is the vast rise in the number of susceptibleindividuals. This is due to a number of factors including new andaggressive immunosuppressive therapies, increased survival in intensivecare, increased numbers of transplant procedures and the greater use ofantibiotics worldwide.

In certain patient groups, fungal infection occurs at high frequency;lung transplant recipients have a frequency of up to 20% colonisationand infection with a fungal organism and fungal infection in allogenichaemopoetic stem cell transplant recipients is as high as 15% (Ribaud etal., 1999, Clin Infect Dis. 28:322-30).

Recently there has been increased awareness of the contribution offungal sensitisation, colonisation, allergy and localised infection inthe exacerbation of existing respiratory diseases. Here fungi have beenimplicated in asthma, COPD, brochiectasis and cystic fibrosis. Allergicbronchopulmonary aspergillosis (ABPA) is a lower respiratory tractcondition caused by fungal colonisation, typically by Apsergillusfumigatus. ABPA can be seen in asthmatics at a rate of 0.7-3.5% andcystic fibrosis at a rate of 7-9%.

Currently there are four classes of antifungal drug are available totreat systemic fungal infections. These are the polyenes (e.g.,amphotericin B), the azoles (e.g., ketoconazole or itraconazole), theechinocandins (e.g., caspofungin) and flucytosine.

The polyenes are the oldest class of antifungal agent being firstintroduced in the 1950's. The exact mode of action remains unclear butpolyenes are only effective against organisms that contain sterols intheir outer membranes. It has been proposed that amphotericin Binteracts with membrane sterols to produce pores allowing leakage ofcytoplasmic components and subsequent cell death.

Azoles work by inhibition of the 14α-demethylase via a cytochromeP450-dependent mechanism. This leads to a depletion of the membranesterol ergosterol and the accumulation of sterol precursors resulting ina plasma membrane with altered fluidity and structure. Echinocandinswork by the inhibition of the cell wall synthetic enzyme β-glucansynthase. This leads to abnormal cell wall formation, osmoticsensitivity and cell lysis.

Flucytosine is a pyrimidine analogue interfering with cellularpyrimidine metabolism as well DNA, RNA and protein synthesis. Howeverwidespread resistance to flucyotosine limits its therapeutic use.

It can be seen that to date the currently available antifungal agentsact primarily against only two cellular targets; membrane sterols(polyenes and azoles) and β-glucan synthase (echinocandins).

Resistance to both azoles and polyenes has been widely reported leavingonly the recently introduced echinocandins to combat invasive fungalinfections. As the use of echinocandins increases, resistance by fungiwill inevitably occur.

The identification of new classes of antifungal agent is required togive the promise of positive therapeutic outcomes to patients.

Pyrrole compounds have also been identified as antifungal agents. WO2009/130481 discloses pyrrole compounds that may be used in theprevention or treatment of fungal disease.

SUMMARY OF THE INVENTION

The present inventors have found that the pyrrole compound2-(1,5-dimethyl-3-phenyl-1H-pyrrol-2-yl)-N-(4-(4-(5-fluoropyrimidin-2-yl)piperazin-1-yl)phenyl)-2-oxoacetamide(the compound of Formula I) is a particularly effective antifungalagent. It shows high potency in enzyme inhibition and fungal inhibitiontests and has good bioavailability and low toxicity. Tests have shownthat this pyrrole compound inhibits the growth of a wide variety offungi, in particular the human pathogenic fungi Aspergillus. Thisparticular compound has been shown to have activity against a widerspectrum of species within the Aspergillus genus than other pyrrolecompounds. Further, the compound has been shown to exhibit increased invivo efficacy when compared to the known antifungal drug Voriconazole,in particular improved efficacy against Scedosporium fungi. The compound2-(1,5-dimethyl-3-phenyl-1H-pyrrol-2-yl)-N-(4-(4-(5-fluoropyrimidin-2-yl)piperazin-1-yl)phenyl)-2-oxoacetamidemay, therefore, be used to effectively treat a wide variety of fungalinfection and disease. These results are described in the internationalpatent application having application number PCT/GB2015/053546, theentirety of which is incorporated by reference herein.

The present inventors have recognised a need for effective formulationsof the pyrrole compound2-(1,5-dimethyl-3-phenyl-1H-pyrrol-2-yl)-N-(4-(4-(5-fluoropyrimidin-2-yl)piperazin-1-yl)phenyl)-2-oxoacetamide.Formulations yielding high bioavailability with minimal side-effects arerequired for the compound to achieve optimal benefit in clinical use.There is further a need for formulations of2-(1,5-dimethyl-3-phenyl-1H-pyrrol-2-yl)-N-(4-(4-(5-fluoropyrimidin-2-yl)piperazin-1-yl)phenyl)-2-oxoacetamidewhich can be easily administered to a patient in need thereof.

Accordingly, the present invention provides a pharmaceutical compositionsuitable for oral administration wherein the composition comprisesspray-dried particles of a compound of Formula I or a pharmaceuticallyacceptable salt thereof

2-(1,5-dimethyl-3-phenyl-1H-pyrrol-2-yl)-N-(4-(4-(5-fluoropyrimidin-2-yl)piperazin-1-yl)phenyl)-2-oxoacetamide

In one aspect, the composition suitable for oral administrationcomprises spray-dried particles of the compound of Formula I. In oneaspect, the compound of Formula I is substantially amorphous. In oneaspect the pharmaceutical composition further comprises one or moreexcipients. In one aspect the composition comprises the excipienthydroxypropyl methyl cellulose acetate succinate (HPMCAS). In oneaspect, the mass ratio of the compound of Formula I to excipient is from1:100 to 1:1, for example from 1:15 to 1:2. In one aspect, thepharmaceutical composition comprises particles which are obtainable byspray-drying from a solution comprising an organic solvent selected fromdichloromethane, methanol, and mixtures thereof. In one aspect, thepharmaceutical composition is in the form of a solid oral dosage form ora liquid oral dosage form. In one aspect, the liquid oral dosage formfurther comprises a pharmaceutically acceptable buffer having a pKa inthe range 6.0 to 8.0. In one aspect, the buffer is from 1 mM to 200 mMphosphate buffer wherein the composition is buffered to about pH 7. Inone aspect, the pharmaceutical further comprises one or morepharmaceutically acceptable binders and/or carriers and/or excipientsand/or diluents and/or adjuvants.

The present invention also provides a pharmaceutical compositionsuitable for parenteral administration comprising (i) a compound ofFormula I or a pharmaceutically acceptable salt thereof, (ii) acyclodextrin or modified cyclodextrin, and (iii) a polyethylene glycol.

In one aspect, the composition suitable for parenteral administrationcomprises from 10 wt % to 40 wt % of a cyclodextrin or modifiedcyclodextrin; and/or from 10 wt % to 40 wt % of a polyethylene glycol.In one aspect, the cyclodextrin or modified cyclodextrin is hydroxypropyl beta cyclodextrin. In one aspect, the polyethylene glycol isPEG300 or PEG400. In one aspect, the composition further comprisespolyvinyl pyrrolidone (Povidone). In one aspect, the compound of FormulaI or the pharmaceutically acceptable salt thereof is present at aconcentration of from 1 mg/mL to 10 mg/mL. In one aspect, thecomposition further comprises one or more pharmaceutically acceptablecarriers and/or excipients and/or diluents and/or adjuvants.

The present invention also provides a pharmaceutical composition asdescribed herein for use in a method of treatment of a human or animalsubject in need thereof, particularly in a method of preventing ortreating fungal infection in the subject. Similarly, the presentinvention provides a method of preventing or treating fungal infectionin a human or animal subject in need thereof, said method comprisingadministering to the human or animal subject a therapeutically effectiveamount of a pharmaceutical composition as described herein; and the useof a pharmaceutical composition as described herein in the manufactureof a medicament for use in the prevention or treatment of fungalinfection in a human or animal subject in need thereof.

The invention further provides a method of producing a pharmaceuticalcomposition comprising a compound of Formula I or a pharmaceuticallyacceptable salt thereof, wherein said method comprises spray-drying asolution of the compound of Formula (I) or salt thereof. In one aspect,the method comprises: (i) dissolving one or more excipients in asolvent; (ii) adding the compound of Formula I to the solution producedin step (i); and (iii) spray drying the solution produced in step (ii).

The invention further provides a pharmaceutical composition suitable fororal administration wherein the composition comprises substantiallyamorphous particles of a compound of Formula I or a pharmaceuticallyacceptable salt thereof.

The compositions of the invention are beneficial as they provideextremely high bioavailability of the compound of Formula I and haveminimal side-effects.

DESCRIPTION OF THE FIGURES

FIG. 1 shows pharmacokinetic data obtained in human trials of aformulation of the compound of Formula I according to the invention asdescribed in Example 5. y-axis: Plasma C_(max) (μg/mL); x-axis: time inhours.

FIG. 2 shows pharmacokinetic data obtained in human trials of aformulation of the compound of Formula I according to the invention asdescribed in Example 5. y-axis: Plasma C_(max) (μg/mL); x-axis: dose ofthe compound of Formula I (mg/kg).

FIG. 3 shows pharmacokinetic data obtained in human trials of aformulation of the compound of Formula I according to the invention asdescribed in Example 5. y-axis: Plasma AUC_(0-∞) (μg·h/mL); x-axis: doseof the compound of Formula I (mg/kg)

DETAILED DESCRIPTION OF THE INVENTION

As used herein, a pharmaceutically acceptable salt is a salt with apharmaceutically acceptable acid or base. Pharmaceutically acceptableacids include both inorganic acids for example hydrochloric, sulphuric,phosphoric, diphosphoric, hydrobromic, hydroiodic or nitric acid andorganic acids for example citric, fumaric, maleic, malic, ascorbic,succinic, tartaric, benzoic, acetic, methanesulphonic, ethanesulphonic,benzenesulphonic, p-toluenesulphonic acid, formic, acetic, propionic,glycolic, lactic, pyruvic, oxalic, salicylic, trichloroacetic, picric,trifluoroacetic, cinnamic, pamoic, malonic, mandelic, bismethylenesalicylic, ethanedisulfonic, gluconic, citraconic, aspartic, stearic,palmitic, EDTA, p-aminobenzoic or glutamic acid, sulfates, nitrates,phosphates, perchlorates, borates, acetates, benzoates,hydroxynaphthoates, glycerophosphates or ketoglutarates. Furtherexamples of pharmaceutically acceptable inorganic or organic acidaddition salts include the pharmaceutically acceptable salts listed inJournal of Pharmaceutical Science, 66, 2 (1977) which are known to theskilled artisan. Pharmaceutically acceptable bases include alkali metal(e.g. sodium or potassium) and alkali earth metal (e.g. calcium ormagnesium) hydroxides and organic bases for example alkyl amines,aralkyl amines and heterocyclic amines, lysine, guanidine,diethanolamine and choline. Also intended as pharmaceutically acceptableacid addition salts are the hydrates which the present compound is ableto form. The acid addition salts may be obtained as the direct productsof compound synthesis. In the alternative, the free base may bedissolved in a suitable solvent containing the appropriate acid, and thesalt isolated by evaporating the solvent or otherwise separating thesalt and solvent. The compound of the active substance may form solvateswith standard low molecular weight solvents using methods known to theskilled artisan.

Unless otherwise stated, all percentages mentioned herein are weightpercentages (wt %).

The invention provides a pharmaceutical composition suitable for oraladministration wherein the composition comprises spray-dried particlesof a compound of Formula I or a pharmaceutically acceptable saltthereof.

2-(1,5-dimethyl-3-phenyl-1H-pyrrol-2-yl)-N-(4-(4-(5-fluoropyrimidin-2-yl)piperazin-1-yl)phenyl)-2-oxoacetamide

Preferably, the composition comprises spray-dried particles of thecompound of Formula I. Most preferably the composition comprises thecompound of Formula I in the form of the free base.

Spray drying is a well-known technique used to produce substantiallyamorphous particles of substances for example pharmaceutically activesubstances e.g. drugs. Spray drying is particularly beneficial forproducing solid particles of thermally-sensitive materials and has theadvantage over other drying techniques that a broadly consistentparticle size is obtained. Any suitable spray drying apparatus can beused to dry the substance, and many spray-dryers are commerciallyavailable. For example, the spray-dryer may be a single effect spraydryer or a multiple effect spray dryer. Multiple effect spray dryers areoften preferred as they can be used to produce uniform particle sizes.Spray drying is described in standard reference texts which are readilyavailable to the skilled person, for example A. S. Mujumdar, Handbook ofIndustrial Drying, CRC Press 2014, which describes spray-dryingtechniques and optimal choices thereof.

Preferably, the pharmaceutical composition suitable for oraladministration comprises a compound of Formula I which is substantiallyamorphous. An amorphous particle is a particle which lacks long-rangecrystallographic ordering. Preferably, the compound of Formula 1 is morethan 50% amorphous, for example more than 70% amorphous, more preferablymore than 90% amorphous, still more preferably more than 95% amorphous,more preferably more than 99% amorphous, for example more than 99.5%amorphous or more than 99.9% amorphous. Substantially amorphousparticles are therefore particles in which the compound of Formula I hasa low crystallinity content, for example less than 50% crystallinity,e.g. less than 30% crystallinity, preferably less than 10%, especiallyless than 5% e.g. less than 1% crystallinity, for example less than 0.5%or less than 0.1% crystallinity. Crystallinity may be measured usingmethods familiar to those skilled in the art. There are many methods oftesting amorphous particles which are known to the skilled person andwhich can be used to determine whether a particle is amorphous orcrystalline. These methods include, but are not limited to powder X-raydiffraction, differential scanning calorimetry, dynamic vapour sorption,isothermal microcalorimetry, inverse gas chromatography, near infra-redspectroscopy and solid-state NMR.

Preferably, the pharmaceutical composition suitable for oraladministration comprises particles of the compound of Formula I whichhave an average particle size of from about 0.5 μm to about 1000 μm,more preferably from about 1 μm to about 500 μm for example from 5 μm to100 μm, for example from about 20 μm to about 50 μm. The term “averageparticle size” refers to the value known as the D50. The term D50 meansthat 50 vol % of the particles have a diameter that is smaller than thisvalue, and 50 vol % of the particles have a diameter that is larger thanthis value. The average particle size may be measured using standardlaser diffraction particle sizing techniques known in the art. Oneexample of an instrument to measure the particle size of the dry powdersis the Mastersizer 2000, manufactured by Malvern Instruments Ltd(Worcestershire, UK).

Preferably, the pharmaceutical composition suitable for oraladministration comprises particles which are obtainable by spray-dryingfrom a solution comprising an organic solvent. Preferably, the organicsolvent is one or more solvents selected from dicholoromethane, acetone,methanol and ethanol. More preferably, the solvent is a mixture of twoor more solvents selected from dicholoromethane, acetone, methanol andethanol. Still more preferably, the solvent is a mixture ofdichloromethane and/or acetone with methanol and/or ethanol. Typically,the ratio of dicholoromethane and/or acetone to methanol and/or ethanolis from 1:1 to 5:1, for example from 2:1 to 4:1 e.g. 3:1. For example,the solvent is often a mixture of dicholoromethane and methanol, whereinthe ratio of dicholoromethane to methanol is from 2:1 to 4:1 for example3:1. Most preferably, the solvent is a 3:1 ratio of dicholoromethane tomethanol. Ratios of solvents can be determined by mass or volume; volumeratios are preferred.

Preferably, the pharmaceutical composition suitable for oraladministration further comprises one or more excipients.Pharmaceutically acceptable excipients known to the skilled personinclude e.g. binding agents, for example syrup, acacia, gelatin,sorbitol, tragacanth, polyvinylpyrrolidone (Povidone), methylcellulose,ethylcellulose, sodium carboxymethylcellulose,hydroxypropylmethylcellulose, sucrose and starch; fillers and carriers,for example corn starch, gelatin, lactose, sucrose, microcrystallinecellulose, kaolin, mannitol, dicalcium phosphate, sodium chloride andalginic acid; and lubricants such as magnesium stearate, sodium stearateand other metallic stearates, glycerol stearate, stearic acid, siliconefluid, talc waxes, oils and colloidal silica. Flavouring agents such aspeppermint, oil of wintergreen, cherry flavouring and the like can alsobe used. It may be desirable to add a colouring agent to make the dosageform readily identifiable. Tablets may also be coated by methods wellknown in the art.

More preferably, the pharmaceutical composition suitable for oraladministration comprises an excipient selected from (i) a cellulose ormodified cellulose for example hypromellose, hydroxypropyl cellulose,hydroxyl propyl methyl cellulose (HPMC), hydroxyl propyl methylcellulose acetate (HPMCA) and hydroxy propyl methyl cellulose acetatesuccinate (HPMCAS) and (ii) a vinylpyrrolidone-vinyl acetate copolymerhaving a mass ratio of vinylpyrrolidone to vinyl acetate in thecopolymer of from 10:1 to 1:10, for example from 5:1 to 1:5 e.g. 3:1 to1:1, for example from 2:1 to 1:1, e.g. 3:2 (which can also be expressedas 6:4), for example Kollidon VA64 or Kollidon VA64 Fine, both availablefrom BASF; Kollidon VA64 is preferred. The pharmaceutical compositionmay comprise a mixture of (i) and (ii). More preferably, thepharmaceutical composition suitable for oral administration comprises acellulose-based excipient according to option (i). Most preferableexcipients include HPMC, HPMCA and HPMCAS; HPMCAS is still morepreferred.

HPMCAS is hydroxypropyl methyl cellulose acetate succinate. The contentof acetyl and succinoyl groups in the polymer can determine theproperties of the HPMCAS. Type L HPMCAS represents polymers with highratio of succinoyl substitution to acetyl substitution; typically 14-18wt % succinoyl content and 5-9 wt % acetyl content. Type M HPMCASrepresents polymers having a lower ratio; typically 10-14 wt % succinoylcontent and 7-11 wt % acetyl content. Type H HPMCAS typically comprises4-8 wt % HPMCAS succinoyl content and 10-14 wt % acetyl content. Type LHPMCAS usually dissolves at around pH≥5.5; type M HPMCAS typicallydissolves around pH≥6.0 and type H HPMCAS typically dissolves aroundpH≥6.8. Usually, the HPMCAS comprises from 12-28 wt % methoxyl and from4-28% hydroxypropoxy. HPMCAS is readily available commercially fromsuppliers such as Shin-Etsu (product AQOAT) and from Ashland(“AquaSolve”). Any suitable HPMCAS known to those skilled in the art canbe used.

Preferably, when an excipient is present in the pharmaceuticalcomposition suitable for oral administration, the mass ratio of thecompound of Formula I to the excipient is from 1:100 to 1:1. Morepreferably, the mass ratio of the compound of Formula 1 to the excipientis from 1:50 to 1:1, for example from 1:25 to 1:1.5 e.g. from 1:15 to1:2, for example 1:10, 1:7, 1:5, 1:4 or 1:3. Most preferably the massratio of the compound of Formula 1 to the excipient is from 1:9 to 1:11,for example 1:10, or is from 1:3 to 1:5, for example 1:4. A mass ratioof the compound of Formula 1 to the excipient of 1:4 can be achieved by,for example, a mass ratio of the compound of Formula 1 to the excipientof 4:16.

The pharmaceutical composition of the invention suitable for oraladministration typically contains from 1 to 50 wt % of the compound ofFormula I; more typically from 4 to 40 wt % for example from 7 to 30 wt%. For example, the pharmaceutical composition may comprise from 5 to 20wt %, for example from 8 to 15 wt %, e.g. from 9 to 11 wt % e.g. 10 wt%. A composition comprising about 10 wt % of the compound of Formula Ican, for example, be produced using a composition comprising only thecompound of Formula I and an excipient for example HPMCAS by using aweight ratio of 1:9 (compound of Formula I:HPMCAS). Alternatively, thepharmaceutical composition may comprise from 10 to 40 wt % of thecompound of Formula I, for example from 15 to 30 wt % e.g. from 18 to 22wt % for example about 20 wt %. A composition comprising 20 wt % of thecompound of Formula I can, for example, be produced using a compositioncomprising only the compound of Formula I and an excipient for exampleHPMCAS by using a weight ratio of 4:16 (compound of Formula I:HPMCAS).

The pharmaceutical composition suitable for oral administration mayfurther comprise one or more pharmaceutically acceptable binders and/orcarriers and/or excipients (i.e. further to excipient(s) above) and/ordiluents and/or adjuvants.

The pharmaceutical composition suitable for oral administration maypreferably be presented as: discrete units such as capsules, sachets ortablets each containing a predetermined amount of the active agent; as apowder or granules; as a solution or a suspension of the active agent inan aqueous liquid or a non-aqueous liquid; or as an oil-in-water liquidemulsion or a water in oil liquid emulsion; or as a bolus etc.Preferably, the pharmaceutical composition suitable for oraladministration is in the form of (i) a solid oral dosage form or (ii) aliquid oral dosage form. More preferably, the pharmaceutical compositionsuitable for oral administration is in the form of a solid oral dosageform.

Solid oral dosage forms include, for example, tablets and capsules.Solid oral forms may contain, together with the active compound,solubilising agents, e.g. cyclodextrins or modified cyclodextrins;diluents, e.g. lactose, dextrose, saccharose, cellulose, corn starch orpotato starch; lubricants, e.g. silica, talc, stearic acid, magnesium orcalcium stearate, and/or polyethylene glycols; binding agents; e.g.starches, arabic gums, gelatin, methylcellulose, carboxymethylcelluloseor polyvinyl pyrrolidone; disaggregating agents, e.g. starch, alginicacid, alginates or sodium starch glycolate; effervescing mixtures;dyestuffs; sweeteners; wetting agents, e.g. lecithin, polysorbates,laurylsulphates; and, in general, non-toxic and pharmacologicallyinactive substances used in pharmaceutical formulations. Suchpharmaceutical preparations may be manufactured in any known manner, forexample, by means of mixing, granulating, tabletting, sugar-coating, orfilm coating processes.

A tablet may be made by compression or moulding, optionally with one ormore accessory ingredients. Compressed tablets may be prepared bycompressing in a suitable machine the active agent in a free flowingform such as a powder or granules, optionally mixed with a binder,lubricant, inert diluent, preservative, surface-active or dispersingagent. Moulded tablets may be made by moulding in a suitable machine amixture of the powdered compound moistened with an inert liquid diluent.The tablets may optionally be coated or scored and may be formulated soas to provide slow or controlled release of the active agent.

Other solid oral dosage forms include lozenges comprising the activeagent in a flavoured base, usually sucrose and acacia or tragacanth andpastilles comprising the active agent in an inert base such as gelatinand glycerin, or sucrose and acacia.

Liquid oral dosage forms include solutions, syrups, emulsions andsuspensions. The solutions may contain solubilising agents e.g.cyclodextrins or modified cyclodextrins. The syrups may contain ascarriers, for example, saccharose or saccharose with glycerine and/ormannitol and/or sorbitol. Liquid oral dosage forms include mouthwashescomprising the active agent in a suitable liquid carrier.

Preferably, when the pharmaceutical composition suitable for oraladministration is in the form of a liquid oral dosage form, the liquidoral dosage form further comprises a pharmaceutically acceptable bufferhaving a pKa in the range 6.0 to 8.0, preferably around pH 7, forexample from pH 6.5 to pH 7.5 e.g. from pH 7.0 to pH 7.5, preferablyaround pH 7.1 to pH 7.3 e.g. around pH 7.2. Any pharmaceuticallyacceptable buffer which is capable of maintaining the pH of the solutionin this range can be used. For example, suitable buffer salts includecitrate (e.g. sodium citrate/citric acid), phosphate (e.g.Na₂HPO₄/NaH₂PO₄) and carbonate (e.g. sodium carbonate/sodiumbicarbonate). Phosphate buffer is preferred. The salt concentration ofthe buffer can be any suitable salt concentration to produce a desiredliquid oral formulation. Generally, the salt concentration in the buffersolution is chosen to maintain the pH of the solution at the desiredvalue, for example at around pH 7 (e.g. pH 7.2). Typical saltconcentrations are from 1 mM to 200 mM, for example from 5 mM to 100 mMe.g. from 10 mM to 50 mM, for example from 20 mM to 40 mM, e.g. around25 mM, around 30 mM, or around 35 mM.

Preferred compositions of the invention suitable for oral administrationtherefore comprise spray-dried particles of a compound of Formula I or apharmaceutically acceptable salt thereof, wherein the compound ofFormula I is substantially amorphous and wherein the composition furthercomprises one or more excipients.

More preferred compositions of the invention suitable for oraladministration comprise spray-dried particles of a compound of FormulaI, wherein the compound of Formula I is substantially amorphous andwherein the composition further comprises the excipient hydroxypropylmethyl cellulose acetate succinate (HPMCAS).

Still more preferred compositions of the invention suitable for oraladministration comprise spray-dried particles of the compound of FormulaI, wherein the compound of Formula I is substantially amorphous andwherein the composition further comprises the excipient HPMCAS, andwherein the mass ratio of the compound of Formula I to the excipient isfrom 1:100 to 1:1, preferably from 1:15 to 1:2.

Yet more preferred compositions of the invention suitable for oraladministration comprise spray-dried particles of the compound of FormulaI, wherein the compound of Formula I is substantially amorphous and isobtainable by spray-drying from a solution comprising an organic solventselected from dichloromethane, methanol, and mixtures thereof, andwherein the composition further comprises the excipient HPMCAS, andwherein the mass ratio of the compound of Formula I to the excipient isfrom 1:15 to 1:2.

Most preferred compositions of the invention suitable for oraladministration comprise spray-dried particles of the compound of FormulaI, wherein the compound of Formula I is substantially amorphous, andwherein the composition comprises 10 wt % of the compound of Formula Iand 90 wt % HPMCAS (i.e. the mass ratio of the compound of Formula I toHPMCAS is 1:9). The compound of formula I is most preferably obtainableby spray-drying from a 3:1 v/v mixture of dichloromethane:methanol.

Similarly, most preferred compositions of the invention suitable fororal administration comprise spray-dried particles of the compound ofFormula I, wherein the compound of Formula I is substantially amorphous,and wherein the composition comprises 20 wt % of the compound of FormulaI and 80 wt % HPMCAS (i.e. the mass ratio of the compound of Formula Ito HPMCAS is 1:4). The compound of formula I is most preferablyobtainable by spray-drying from a 3:1 v/v mixture ofdichloromethane:methanol.

The invention also provides a pharmaceutical composition suitable fororal administration wherein the composition comprises substantiallyamorphous particles of a compound of Formula I or a pharmaceuticallyacceptable salt thereof. The composition is as described herein.

More preferably, the invention provides a pharmaceutical compositionsuitable for oral administration comprising the compound of Formula Iwherein the compound of Formula I is substantially amorphous and thecomposition further comprises HPMCAS, wherein the mass ratio of thecompound of Formula I to the HPMCAS is from about 1:3 to 1:5, forexample 1:4, or from about 1:8 to 1:10, for example about 1:9.Therefore, the invention provides a pharmaceutical composition suitablefor oral administration comprising the compound of Formula I and HPMCAS,wherein the compound of Formula I is substantially amorphous and thecomposition comprises 10 wt % of the compound of Formula I and 90 wt %HPMCAS. The invention also provides a pharmaceutical compositionsuitable for oral administration comprising the compound of Formula Iand HPMCAS, wherein the compound of Formula I is substantially amorphousand the composition comprises 20 wt % of the compound of Formula I and80 wt % HPMCAS.

The invention also provides a method of producing a pharmaceuticalcomposition comprising a compound of Formula I or a pharmaceuticallyacceptable salt thereof

2-(1,5-dimethyl-3-phenyl-1H-pyrrol-2-yl)-N-(4-(4-(5-fluoropyrimidin-2-yl)piperazin-1-yl)phenyl)-2-oxoacetamide

wherein said method comprises spray-drying a solution of the compound ofFormula (I) or salt thereof.

Any suitable spray-drying technique can be used. Spray-drying isdescribed above.

Preferably, the method of producing a pharmaceutical compositionsuitable for oral administration comprises the steps of:

i) dissolving one or more excipients in a solvent;

ii) adding the compound of Formula I to the solution produced in step(i); and

iii) spray drying the solution produced in step (ii).

Preferably, one or more of the one or more excipients is an excipientdescribed herein. More preferably, the one or more excipients is (i) acellulose or modified cellulose as described herein or (ii) avinylpyrrolidone-vinyl acetate copolymer as described herein, or amixture of (i) and (ii). Most preferably, the excipient is HPMCAS asdescribed herein.

Preferably, the solvent is an organic solvent as described herein. Morepreferably, the solvent is a mixture of dichloromethane and/or acetonewith methanol and/or ethanol. Still more preferably, the solvent is amixture of dicholoromethane and methanol, wherein the ratio ofdicholoromethane to methanol is from 2:1 to 4:1 for example 3:1. Mostpreferably, the solvent is a 3:1 ratio of dicholoromethane to methanol.

For example, the invention therefore provides a method of producing apharmaceutical composition suitable for oral administration as describedherein wherein:

-   -   the excipient is hydroxypropyl methyl cellulose acetate        succinate (HPMCAS);    -   the solvent is a mixture of dichloromethane and methanol,        wherein the volume ratio of dichloromethane to methanol is from        5:1 to 1:1;    -   the concentration of the excipient in the solvent is from 5% to        20% w/v; and    -   the compound of Formula I is added to the solution of the        excipient in the solvent to give a concentration of 0.5% to 10%        by mass.        More preferably, the invention provides a method of producing a        pharmaceutical composition suitable for oral administration as        described herein wherein:    -   the excipient is hydroxypropyl methyl cellulose acetate        succinate (HPMCAS);    -   the solvent is a mixture of dichloromethane and methanol,        wherein the volume ratio of dichloromethane to methanol is from        4:1 to 2:1;    -   the concentration of the excipient in the solvent is from 7% to        18% w/v; and    -   the compound of Formula I is added to the solution of the        excipient in the solvent to give a concentration of 0.5% to 6%        by mass.        Still more preferably, the invention provides a method of        producing a pharmaceutical composition suitable for oral        administration as described herein wherein:    -   the excipient is hydroxypropyl methyl cellulose acetate        succinate (HPMCAS);    -   the solvent is a mixture of dichloromethane and methanol,        wherein the volume ratio of dichloromethane to methanol is about        3:1; and    -   (i) the concentration of the excipient in the solvent is from        about 7 wt % to about 11 wt %; and the compound of Formula I is        added to the solution of the excipient in the solvent to give a        concentration of about 0.5% to about 2% by mass; or    -   (ii) the concentration of the excipient in the solvent is from        about 12% to about 18%; and the compound of Formula I is added        to the solution of the excipient in the solvent to give a        concentration of about 3% to about 5% by mass        Most preferably, the invention provides a method of        pharmaceutical composition suitable for oral administration        comprising    -   i) dissolving from about 7 wt % to about 11 wt % (e.g. about 9        wt %) of HPMCAS in a solvent wherein the solvent is a 3:1 v/v        mixture of dichloromethane:methanol;    -   ii) adding the compound of Formula I to the solution produced in        step (i) to yield a solution wherein the concentration of the        compound of Formula I is about 0.5% to about 2% by mass (e.g.        about 1% by mass); and    -   iii) spray drying the solution produced in step (ii).        Similarly, the invention provides a method of pharmaceutical        composition suitable for oral administration comprising    -   i) dissolving from about 12 wt % to about 18 wt % (e.g. about 16        wt %) of HPMCAS in a solvent wherein the solvent is a 3:1 v/v        mixture of dichloromethane:methanol;    -   ii) adding the compound of Formula I to the solution produced in        step (i) to yield a solution wherein the concentration of the        compound of Formula I is about 3% to about 5% by mass (e.g.        about 4% by mass); and    -   iii) spray drying the solution produced in step (ii).

The invention also provides a pharmaceutical composition suitable forparenteral administration comprising (i) a compound of Formula I or apharmaceutically acceptable salt thereof,

2-(1,5-dimethyl-3-phenyl-1H-pyrrol-2-yl)-N-(4-(4-(5-fluoropyrimidin-2-yl)piperazin-1-yl)phenyl)-2-oxoacetamide

(ii) cyclodextrin or modified cyclodextrin, and (iii) polyethyleneglycol.

Pharmaceutically acceptable salts are described herein. Preferably, thepharmaceutical composition suitable for parenteral administrationcomprises a compound of Formula I. More preferably, the pharmaceuticalcomposition suitable for parenteral administration comprises a compoundof Formula I in the form of the free base.

Preferably, cyclodextrin is present in an amount of from 10 wt % to 40wt % with reference to the pharmaceutical composition. Typical amountsof cyclodextrin present in the composition are from 20 wt % to 30 wt %for example about 25 wt %.

Preferably, polyethylene glycol is present in an amount of from 10 wt %to 40 wt % with reference to the pharmaceutical composition. Typicalamounts of polyethylene glycol present in the composition are from 20 wt% to 30 wt % for example about 25 wt %.

The pharmaceutical composition suitable for parenteral administrationmay comprise any suitable cyclodextrin or mixture thereof. Typicalcyclodextrins contain a number of glucose monomers ranging from six toeight units in a ring, creating a cone shape. Cyclodextrins are oftendenoted as a (alpha)-cyclodextrin (comprising a 6-membered sugar ringmolecule); 13 (beta)-cyclodextrin (comprising a 7-membered sugar ringmolecule) or y (gamma)-cyclodextrin (comprising an 8-membered sugar ringmolecule). Because cyclodextrins are hydrophobic inside and hydrophilicoutside, they can form complexes with hydrophobic compounds. Thus theycan enhance the solubility and bioavailability of such compounds.Alpha-, beta-, and gamma-cyclodextrin are all generally recognized assafe by the FDA. Cyclodextrins (CDs) can be modified in various wayswhilst still be suitable for use in the compositions of the invention.For example, known modified cyclodextrins include Hydroxyethyl-β-CD(HE-β-CD), Hydroxypropyl-β-CD (HP-β-CD), Sulfobutylether-β-CD(SBE-β-CD), Methyl-β-CD (M-β-CD), Dimethyl-β-CD (DM-β-CD/DIMEB),Randomly dimethylated-β-CD (RDM-β-CD), Randomly methylated-β-CD(RM-β-CD/RAMEB), Carboxymethyl-β-CD (CM-β-CD), Carboxymethyl ethyl-β-CD(CME-β-CD), Diethyl-β-CD (DE-β-CD), Tri-O-methyl-β-CD (TRIMER),Tri-O-ethyl-β-CD (TE-β-CD), Tri-O-butyryl-β-CD (TB-β-CD),Tri-O-valeryl-β-CD (TV-β-CD), Di-O-hexanoyl-β-CD (DH-β-CD),Glucosyl-β-CD G1-β-CD Maltosyl-β-CD (G2-β-CD), and2-hydroxy-3-trimethyl-ammoniopropyl-β-CD (HTMAPCD). Cyclodextrins andtheir uses in pharmaceutical formulations are disclosed in standardreference texts, for example Fromming and Szejtli, Cyclodextrins inPharmacy, Springer, 1993, which describes the advantages of specificcyclodextrins in pharmaceutical formulations.

Preferably, the cyclodextrin is selected from hydroxy propyl betacyclodextrin and sulfobutyl ether beta-cyclodextrin (Captisol) andmixtures thereof. Hydroxy propyl beta cyclodextrin is preferred.

The pharmaceutical composition suitable for parenteral administrationmay comprise any suitable polyethylene glycol or mixture thereof. Forexample, the composition may comprise any polyethylene glycol approvedfor intraveneous use. Polyethylene glycols which can be used includePEG200-PEG500, for example PEG300 and/or PEG400. PEG 300 and PEG 400 arepreferred, and PEG400 is most preferred.

As the skilled person will appreciate, the number following the term“PEG” (e.g. 300 in “PEG300”) refers to the average molecular weights ofthe PEG molecule. PEG400 thus typically comprises approximately 9ethylene glycol units in each polymer molecule, and PEG 300 typicallycomprises 7 ethylene glycol units in each polymer molecule. However, asthe skilled person will appreciate, many commercially available PEGs arepolydisperse. In general terms, the molecular weight distribution can becharacterized statistically in terms of its weight average molecularweight (Mw) and its number average molecular weight (Mn), the ratio ofwhich is often referred to as the polydispersity index (Mw/Mn). Both Mwand Mn can be measured by conventional techniques, for example, by massspectrometry.

Preferably, the pharmaceutical composition suitable for parenteraladministration comprises one or more dispersing agents for example lowmolecular weight povidone (polyvinyl pyrrolidone). Preferably, thepovidone is endotoxin-free. Povidones are available from commercialsuppliers for example Ashland (Plasdone). Preferably the povidone has aK-value of from 5 to 20 for example from 10 to 18. For example, thePovidone may have a nominal molecular weight of about 4000 and a K valueof from about 10 to about 14. Alternatively the povidone may have anominal molecular weight of about 10000 and a K value of from about 15to about 18. The K value is a function of the average degree ofpolymerisation and the intrinsic viscosity of the polymer and can becalculated from the kinematic viscositiy of an aqueous solution of thepolymer. Preferably, the povidone has a T_(g) (glass transitiontemperature) of from about 110° C. to about 130° C. e.g. from about 120°C. to about 126° C. The pharmaceutical composition may comprise amixture of two or more povidones.

Preferably, when the pharmaceutical composition suitable for parenteraladministration comprises a povidone or mixture thereof, the povidone ormixture thereof is present in an amount of from 0.1 to 5 wt %, morepreferably from 0.5 to 2 wt %, still more preferably about 1 wt %relative to the total mass of the composition.

Preferably, the compound of Formula I is present in the pharmaceuticalcomposition suitable for parenteral administration at a concentration offrom 1 mg/mL to 10 mg/mL. More preferably the concentration of thecompound of Formula I in the pharmaceutical composition suitable forparenteral administration is from 2 to 7 mg/mL for example from 3 to 5mg/mL e.g. 4 mg/mL.

The pharmaceutical composition suitable for parenteral administrationmay further comprise one or more pharmaceutically acceptable carriersand/or excipients and/or diluents and/or adjuvants. For example, thecomposition may contain as carrier, for example, sterile water, or maybe in the form of a sterile, aqueous, isotonic saline solutions.

Preferably, the pharmaceutical composition suitable for parenteraladministration is adjusted to a final pH of from about pH 4 to about pH8.

More preferably, the pharmaceutical composition suitable for parenteraladministration is adjusted to a final pH of from about pH 4 to about pH6, for example from about pH 4.5 to about pH 5.5, e.g. about pH 5, forexample pH 5.0. The pH of the pharmaceutical composition may be adjustedusing any pharmaceutically acceptable acid or base. Phosphoric acid ispreferred.

Preferred pharmaceutical compositions are sterile and pyrogen free.

Preferred compositions of the invention suitable for parenteraladministration therefore comprise:

-   -   from 10 wt % to 40 wt % cyclodextrin or modified cyclodextrin;    -   from 10 wt % to 40 wt % polyethylene glycol; and    -   a dispersing agent for example povidone.

For example, compositions of the invention suitable for parenteraladministration may comprise:

-   -   from 1 to 10 mg/mL of the compound of Formula I or a        pharmaceutically acceptable salt thereof;    -   from 10 wt % to 40 wt % hydroxyl propyl beta cyclodextrin;    -   from 10 wt % to 40 wt % PEG300 or PEG400; and    -   a dispersing agent for example povidone, wherein the povidone is        as described herein; and        wherein the pH of the composition is adjusted to from about pH 4        to about pH 8.

More preferred compositions of the invention suitable for parenteraladministration comprise:

-   -   from 1 to 10 mg/mL of the compound of Formula I or a        pharmaceutically acceptable salt thereof;    -   from 10 wt % to 40 wt % hydroxyl propyl beta cyclodextrin;    -   from 10 wt % to 40 wt % PEG300 or PEG400; and    -   a dispersing agent for example povidone, wherein the povidone is        as described herein; and        wherein the pH of the composition is adjusted to from about pH 4        to about pH 6.

Still more preferred compositions of the invention suitable forparenteral administration comprise:

-   -   from 3 to 5 mg/mL of the compound of Formula I;    -   from 20 wt % to 30 wt % hydroxyl propyl beta cyclodextrin;    -   from 20 wt % to 30 wt % PEG300 or PEG400, preferably PEG400; and    -   from 0.1 to 5 wt % povidone; and        wherein the pH of the composition is adjusted to from about pH        4.5 to about pH 5.5.

Most preferred compositions of the invention suitable for parenteraladministration comprise:

-   -   4 mg/mL (relative to the final volume of the composition) of the        compound of Formula I    -   wt % hydroxy propyl beta cyclodextrin;    -   wt % PEG400;    -   1 wt % polyvinyl pyrrolidone (Povidone);    -   phosphoric acid in sufficient amount to adjust the pH of the        pharmaceutical composition to pH 5.0; and    -   water to 100%.

As described herein, a pharmaceutical composition of the invention canfurther comprise one or more adjuvants for example a local anaesthetic,preservative or buffering agent.

Pharmaceutically acceptable binders include solution binders and drybinders. Solution binders are dissolved in a solvent (for example wateror alcohol can be used in wet granulation processes). Examples includegelatin, cellulose, cellulose derivatives, polyvinylpyrrolidone, starch,sucrose and polyethylene glycol. Dry binders are added to the powderblend, either after a wet granulation step, or as part of a directpowder compression (DC) formula. Examples include cellulose, methylcellulose, polyvinylpyrrolidone and polyethylene glycol.

Pharmaceutical carriers include liposomes, nanospheres, micelles,protein-DNA complexes, nanogels and natural solvents for example aqueousand non-aqueous solutions.

Other substances useful as excipients, diluents or carriers includeacacia, alginate, alginic acid, aluminum acetate, benzyl alcohol, butylparaben, butylated hydroxy toluene, citric acid, calcium carbonate,candelilla wax, croscarmellose sodium, confectioner's sugar, colloidalsilicone dioxide, cellulose, calcium phosphate, carnuba wax, cornstarch, carboxymethylcellulose calcium, calcium stearate, calciumdisodium EDTA, copolyvidone, hydrogenated castor oil, calcium hydrogenphosphate dehydrate, cetylpyridine chloride, cysteine HCl,crosspovidone, disodium hydrogen phosphate, dimethicone, sodiumerythrosine, ethyl cellulose, gelatin, glyceryl monooleate, glycerine,glycine, glyceryl monostearate, glyceryl behenate, hydroxy propylcellulose, hydroxyl propyl methyl cellulose, hypromellose, HPMCpthalate, lactose, magnesium stearate, mannitol, methyl cellulose,magnesium carbonate, mineral oil, magnesium oxide, methyl paraben,povidone, polysorbate 80, polyethylene oxide, polaxamer 407 or 188,potassium bicarbonate, potassium sorbate, potato starch, phosphoricacid, polyoxy stearate, sodium starch glycolate, sodium crossmellose,sodium lauryl sulphate, starch, silicon dioxide, sodium benzoate,stearic acid, sucrose, sorbic acid, sodium carbonate, saccharin sodium,sodium alginate, silica gel, sorbiton monooleate, sodium stearylfumarate, sodium chloride, sodium metabisulfite, sodium citratedehydrate, sodium carboxy methyl cellulose, succinic acid, sodiumpropionate, titanium dioxide, talc, triacetin and triethyl citrate.

A pharmaceutical composition as described herein may comprise particlesof the compound of Formula I wherein the average particle size (asdescribed herein) has undergone particle size reduction by micronisationor nanonisation technologies.

The compositions of the invention are particularly advantageous as theyprovide the compound of Formula I with increased bioavailability. Asused herein, bioavailability is defined such that a drug for example thecompound of Formula I, when administered intraveneously, has abioavailability of 100%. The pharmaceutical compositions of theinvention which are suitable for oral administration are particularlybeneficial as they can provide the compound of Formula I with highbioavailability. Preferably, the composition of the invention providesthe compound of Formula I with a bioavailability of at least 50%, morepreferably at least 70% for example at least 80%, still more preferablyat least 90% for example at least 95%. As the skilled person willappreciate, bioavailability can be determined by numerous factorsincluding the nature of the subject to which the composition isadministered (age, weight, sex etc). Therefore, a negative result in onesubject group is not determinative.

Bioavailability can be determined by pharmacokinetic (PK) studies inwhich plasma drug concentration is determined as a funcation of timeafter both intravenous (IV) and extravascular (e.g., oral)administration. The absolute bioavailability (F_(abs)) is the dose(D)-corrected area under curve (AUC) (non-intravenous) divided by AUC(intravenous). As used herein, F_(abs) for a drug administered by theoral route (PO) is calculated using:

$F_{abs} = {100\frac{{AUC}_{PO} \cdot D_{IV}}{{AUC}_{IV} \cdot D_{PO}}}$

The compositions of the invention are useful in treating medicalconditions in a human or animal subject in need thereof.

Accordingly, the invention provides a pharmaceutical composition asdescribed herein for use in a method of treatment of a human or animalsubject in need thereof. Preferably, the invention provides apharmaceutical composition as described herein for use in the treatmentof a human or animal subject in need thereof, wherein the treatmentcomprises prevention of or treatment of fungal infection in the subject.

The invention also provides a method of preventing or treating fungalinfection in a human or animal subject in need thereof, said methodcomprising administering to the human or animal subject atherapeutically effective amount of a pharmaceutical composition asdescribed herein.

The invention also provides the use of a pharmaceutical composition asdescribed herein in the manufacture of a medicament for use in theprevention or treatment of fungal infection in a human or animal subjectin need thereof. A pharmaceutical composition of the invention may beused in a method of treating a human or animal subject wherein thetreatment comprises administration of the composition in combinationwith a further antifungal agent as described herein.

A therapeutically effective amount of a composition of the invention maybe administered to a patient in need thereof. For example, thecomposition is typically administered in an amount such as to provide tothe subject a daily dose of the compound of Formula I of up to 200 mg,e.g. up to 100 mg or up to 50 mg per kg of body weight, for example from0.001 to 200 or 0.001 to 50 mg per kg of body weight, according to (forexample) the age, weight and conditions of the subject to be treated,the type and severity of the disease and the frequency and route ofadministration. Preferably, daily dosage levels are up to 200 mg, e.g.up to 150 mg, up to 100 mg, up to 50 mg or up to 40 mg per kg of bodyweight. Daily dosage levels are for example at least 1 mg, at least 2 mgor at least 5 mg per kg of body weight. In one embodiment the dailydosage level is from 0.05 mg to 2 g, preferably from 0.1 mg to 10 mg.The appropriate dosage level can be readily determined by the skilledphysician.

Where a composition of the invention is administered together with asecond antifungal agent, the second antifungal agent is typicallyadministered at or below the standard dose used for that drug. In thismanner, known antifungal agents may be administered in lower doses thanare currently used, resulting in a reduction in toxic effects.

The composition of the invention is useful in the treatment of orprevention of fungal disease. Preferably, the fungal disease comprisesan infection by a fungus, for example an Ascomycete. Preferably, thefungal disease comprises an infection by an organism selected from thegenera Absidia; Acremonium; Alternaria; Aspergillus; Bipolaris;Blastomyces; Blumeria; Cladosporium; Coccidioides; Colletotrichium;Curvularia; Encephalitozoon; Epicoccum; Epidermophyton; Exophiala;Exserohilum; Fusarium; Histoplasma; Leptosphaeria; Microsporum;Mycosphaerella; Neurospora, Paecilomyces; Penicillium; Phytophthora;Plasmopara; Pneumocystis; Pyricularia; Pythium; Puccinia; Rhizoctonia;Rhizomucor; Scedosporium; Scopulariopsis; Trichophyton; Trichosporon;and Ustilago.

Preferably, the fungal disease comprises an infection by an organism ofthe genus Aspergillus, Scedosporium or Fusarium, for instance, thefungal disease comprises an infection by an organism of the genusAspergillus or Scedosporium, in particular Aspergillus. In oneembodiment, the fungal disease comprises an infection by an organism ofthe genus Aspergillus. In another embodiment, the fungal diseasecomprises an infection by an organism of the genus Scedosporium.

Preferably, the fungal disease comprises an infection by an organismselected from the species Absidia corymbifera; Acremonium spp;Alternaria alternata; Aspergillus flavus; Aspergillus fumigatus;Aspergillus nidulans; Aspergillus niger; Aspergillus parasiticus;Aspergillus terreus; Bipolaris spp; Blastomyces dermatitides; Blumeriagraminis; Cladosporium cladosporoides; Cladosporium herbarium;Coccidioides immitis; Coccidioides posadasii; Curvularia lunata;Colletotrichium trifolii; Encephalitozoon cuniculi; Epicoccum nigrum;Epidermophyton floccosum; Exophiala spp; Exserohilum rostratum; Fusariumgraminarium; Fusarium solani; Fusarium sporotrichoides; Histoplasmacapsulatum; Leptosphaeria nodorum; Microsporum canis; Mycosphaerellagraminicola; Paecilomyces lilanicus; Paecilomyces varioti; Penicilliumchrysogenum; Phytophthora capsici; Phytophthora infestans; Plasmoparaviticola; Pneumocystis jiroveci; Puccinia coronata; Puccinia graminis;Pyricularia oryzae; Pythium ultimum; Rhizoctonia solani; Rhizomucor spp;Rhizopus spp; Scedosporium apiospermum; Scedosporium prolificans;Scedosporium species d; Scopulariopsis brevicaulis; Trichophytonmentagrophytes; Trichophyton interdigitale; Trichophyton rubrum;Trichosporon asahii; Trichosporon beigelii; and Ustilago maydis.

Preferably, the fungal disease comprises an infection by A. fumigatus,A. flavus, A. terreus, A. niger, A. lentulus, S. apiospermum, S.prolificans, or S. species d. Particularly, the fungal disease comprisesan infection by A. fumigatus, A. flavus, A. terreus or A. niger. In oneembodiment, the fungal disease comprises an infection by S. prolificans.

Examples of fungal diseases which can be prevented or treated using acomposition of the invention include both systemic and superficialinfections. The fungal diseases include invasive fungal diseases causedby Aspergillus species for example aspergillosis, but also local formsof these infections. For instance, the fungal diseases include invasivefungal diseases caused by Aspergillus species for example aspergillosis,but also local forms of these infections. Compositions of the inventionis particularly useful against diseases caused by Aspergillus species,for which a fungicidal drug is required which has lower toxicity thanamphotericin. The invention also provides for the treatment ofdermatological infections.

The pharmaceutical composition of the invention is, in one embodiment,for use in the prevention or treatment of a disease caused byAspergillus species. The diseases caused by Aspergillus species includediseases caused by A. fumigatus, A. flavus, A. terreus and A. niger.

Examples of systemic infections which might be prevented or treatedusing a pharmaceutical composition of the invention include: pulmonaryaspergillosis, e.g. in immunosuppressed patients for example bone marrowrecipients or AIDS patients; systemic aspergillosis; rhinocerebralmucomycosis; blastomycosis; histoplasmosis; coccidiomycosis;paracoccidiomycosis; lobomycosis; sporotrichosis; chromoblastomycosis;phaeohyphomycosis; and disseminated sporotrichosis.

Examples of superficial infections, which can be prevented or treatedusing a pharmaceutical composition of the invention include: ring worm;athlete's foot; and tinea unguium (nail infection).

Examples of diseases or conditions which are caused by fungi or wherefungi exacerbate an allergic response, and which can be prevented ortreated using a pharmaceutical composition of the invention includeallergic bronchopulmonary aspergillosiis (ABPA); asthma, Severe asthmawith Fungal Sensitisation (SAFS), fungal colonization of cysticfibrosis, rhinosinusitis and sinusitis. For instance, the disease may becaused by a fungal sensitisation, or the disease may be AllergicBronchopulmonary Aspergillosis (ABPA) or asthma.

The pharmaceutical compositions described herein can be administered incombination with a second antifungal agent. Preferably, thepharmaceutical composition is administered separately from orsuccessively with the second antifungal agent. For example, thecomposition of the invention and the second antifungal agent may beprovided as a kit. The kit may thus comprise a composition of theinvention and a second antifungal agent.

The second antifungal agent can be any suitable antifungal agent thatthe skilled person would judge to be useful in the circumstances. Forexample, any of the conditions described herein can be treated in hismanner.

Particularly suitable classes of antifungal agents include azoles,polyenes, purine nucleotide inhibitors, pyrimidine nucleotideinhibitors, mannan inhibitors, protein elongation factor inhibitors,chitin synthase inhibitors, Beta-glucan synthase inhibitors,echinocandins, allylamines, anti-HSP90 antibodies,bactericidal/permeability inducing protein products and polyoxins. Othersuitable antifungal agents which do not fall within the classes aboveinclude the compounds 5-fluoro-1,3-dihydro-1-hydroxy-2,1-benzoxaborale(AN269), 5-chloro-1,3-dihydro-1-hydroxy-2,1-benzoxaborale (AN2718) andicofungipen.

For instance, the second antifungal agent may be selected from the groupconsisting of azoles, polyenes, purine nucleotide inhibitors, pyrimidinenucleotide inhibitors, mannan inhibitors, protein elongation factorinhibitors, echinocandins, allylamines, anti-HSP90 antibodies,bactericidal/permeability inducing protein products or polyoxins, or oneof the compounds 5-fluoro-1,3-dihydro-1-hydroxy-2,1-benzoxaborale(AN269), 5-chloro-1,3-dihydro-1-hydroxy-2,1-benzoxaborale (AN2718),icofungipen, VT116 or SCY078.

VT116 is 2-Pyridineethanol,α-(2,4-difluorophenyl)-β,β-difluoro-α-(1H-tetrazol-1-ylmethyl)-5-[4-(2,2,2-trifluoroethoxy)phenyl]-,(αR)-,

and SCY078 078 (aka MK-3118) is a semi-synthetic derivative ofenfumafungin, 4H-1,4a-Propano-2H-phenanthro[1,2-c]pyran-7-carboxylicacid,15-[(2R)-2-amino-2,3,3-trimethylbutoxy]-8-[(1R)-1,2-dimethylpropyl]-1,6,6a,7,8,9,10,10a,10b,11,12,12a-dodecahydro-1,6a,8,10a-tetramethyl-14-[5-(4-pyridinyl)-1H-1,2,4-triazol-1-yl]-,(1S,4aR,6aS,7R,8R,10aR,10bR,12aR,14R,15R):

Preferred azoles are clotrimazole, econazole, bifonazole, butoconazole,fenticonazole, fluconazole, isoconazole, itraconazole, ketoconazole,miconazole, oxiconazole, sertaconazole, sulconazole, tioconazole,isavuconazole, ravuconazole, posaconazole, terconazole and voriconazole,luliconazole. Preferred echinocandins are anidulafungin, caspofunginmicafungin and biafungin. Preferred allylamines are terbinafine,butenafine, amorolfine and naftifine. Preferred polyenes areamphotericin B and nystatin. A preferred example of a purine orpyrimidine nucleotide inhibitor is flucytosine. A preferred mannaninhibitor is pradamicin. A preferred protein elongation factor inhibitoris sordarin and analogues thereof. A preferred polyoxin is nikkomycin Z.

Particularly preferred second antifungal agents are caspofungin,micafungin, anidulofungin, amphotericin B, voriconazole, posaconazole,isavuconazole, fluconazole and itraconazole.

Synthesis

The compound of Formula I is2-(1,5-dimethyl-3-phenyl-1H-pyrrol-2-yl)-N-(4-(4-(5-fluoropyrimidin-2-yl)piperazin-1-yl)phenyl)-2-oxoacetamide,or a pharmaceutically acceptable salt thereof.

2-(1,5-dimethyl-3-phenyl-1H-pyrrol-2-yl)-N-(4-(4-(5-fluoropyrimidin-2-yl)piperazin-1-yl)phenyl)-2-oxoacetamide

One synthetic route to the compound of Formula I is described herein. Ingeneral terms, the compound of Formula I may be synthesised by reactinga compound of formula (II), with a compound of formula (III). Typicallythe reaction takes place in the presence of an organic solvent and abase. Preferably the solvent is dichloromethane or tetrahydrofuran andthe base is triethylamine or pyridine. Typically the reaction is carriedout at 0° C. initially while the reagents are added and then stirred atroom temperature until the reaction is complete. The compound of formula(III) is typically available from commercial sources or can be preparedby known methods.

The compound of formula (II) may be prepared by reacting a compound offormula (IV), with preferably oxalyl chloride. Typically the reactiontakes place in an organic solvent. Preferably, the solvent isdichloromethane. Typically, the reaction is carried out at 0° C.initially while the reagents are added and then stirred at roomtemperature until the reaction is complete.

All of the starting materials referred to in the reactions describedabove are available from commercial sources or can be prepared byanalogy with known methods.

The following examples illustrate the invention but are not intended tolimit the scope of the invention. In this regard, it is important tounderstand that the particular assays used in the Examples section aredesigned only to provide an indication of anti-fungal activity. Thereare many assays available to determine such activity, and a negativeresult in any one particular assay is therefore not determinative.

EXAMPLES Example 1: Synthesis of the Compounds of Formula I(2-(1,5-dimethyl-3-phenyl-1H-pyrrol-2-yl)-N-(4-(4-(5-fluoropyrimidin-2-yl)piperazin-1-yl)phenyl)-2-oxoacetamide)

The synthesis of the compound of Formula I is described in theinternational patent application having application numberPCT/GB2015/053546. Information relating to the synthesis of the compoundof Formula I is incorporated by reference. The following Example isreproduced from that patent application.

The synthetic scheme below provides a method of synthesis of:

2-Hydroxyimino-3-oxo-3-phenyl propionic acid ethyl ester (A)

A solution of sodium nitrite (1.07 Kg, 45.62 mol) in water (4 L) wasadded slowly to a solution of ethyl benzoyl acetate (2 Kg, 10.41 mol) inglacial acetic acid (6 L), at 0-10° C. over a period of 2 h. The productstarted precipitating during the course of addition. The reaction masswas warmed to room temperature and stirred for a further 1 h. Water (2.5L) was added and the mixture stirred for a further 1 h. Filtered undersuction, washed with water (2 L). The solid was dissolved in chloroform(8 L) and washed with water (2×500 mL), brine solution (2×500 mL), driedover anhydrous sodium sulfate and concentrated in vacuo to dryness toafford 2.0 Kg (86%) of 2-hydroxyimino-3-oxo-3-phenyl propionic acidethyl ester A as a white solid. [TLC system: Ethyl acetate:Pet ether(3:7); R_(f) value: 0.28].

5-Methyl-3-phenyl-1H-pyrrole-2,4 dicarboxylic acid diethyl ester (B)

A mixture of ethyl acetoacetate (329 g, 2.53 mol), zinc dust (443 g,6.78 mol) and anhydrous sodium acetate (463 g, 5.65 mol) in glacialacetic acid (800 mL) were heated to 60° C. A solution of A (500 g, 2.26mol) in glacial acetic acid (1.5 L) was added in three portions undervigorous stirring over a period of ˜1 h. The temperature shot up toabout 93° C. during the addition. The reaction mixture was maintained at60-75° C. for 3 h. Additional zinc dust (221 g, 3.39 mol) was added tothe reaction mass over 15 min and the mixture was stirred at 60-75° C.for 1 h, cooled to room temperature and filtered the solids. Thefiltrate was evaporated in vacuo and the residue was co-distilled withtoluene (2×500 mL). Water (5 L) and ethyl acetate (1 L) were added tothe residue and stirred till two clear layers were obtained. The organiclayer washed successively with water (2×500 mL), saturated bicarbonatesolution (2×500 mL), brine (2×500 mL) dried over anhydrous sodiumsulfate and concentrated to give 360 g of crude gummy product. This wasstirred with a mixture of dichloromethane in pet ether (200 mL: 1200 mL;1:6) at room temperature for 15 min, filtered and washed with pet ether(100 mL) to afford 250 g (36%) of 5-methyl-3-phenyl-1H-pyrrole-2,4dicarboxylic acid diethyl ester B as off-white solid. [TLC system: ethylacetate:Pet ether (3:7); R_(f) value: 0.45]. Similarly 1.5 Kg (500 g×3)of A was converted to 500 g [245 g (36%)+255 g (37%)+250 g (36%)] of Bin three batches.

1, 5-Dimethyl-3-phenyl-1H-pyrrole-2,4-dicarboxylic acid diethyl ester(C)

A solution of B (1 Kg, 3.322 mol) in dry tetrahydrofuran (4 L) was addedto slurry of sodium hydride (60% w/w; 254 g, 6.644 mol) in drytetrahydrofuran (4 L) at 0° C. over 1 h. The reaction mass was warmed toroom temperature and stirred for 1 h and again cooled to 0° C. Methyliodide (517 mL; 8.305 mol) was added over ½ h and the reaction mixturestirred at room temperature for 18 h. Quenched with ice-water (100 mL)and 1N hydrochloric acid (2 L) was added. The organic layer wasseparated and the aqueous layer was extracted with dichloromethane(2×500 mL). The combined organic layers were washed successively withbrine (2×200 mL), dried over anhydrous sodium sulfate and concentratedto dryness to afford 950 g (91%) of 1, 5-dimethyl-3-phenyl-1H-pyrrole-2,4-dicarboxylic acid diethyl ester C as a yellow solid [TLC system: ethylacetate:Pet ether (3:7); R_(f) value: 0.56].

1, 5-Dimethyl-3-phenyl-1H-pyrrole-2,4-dicarboxylic acid (D)

A solution of sodium hydroxide (1.21 Kg, 30.25 mol) in water (3.6 L) wasadded to a solution of C (950 g, 3.025 mol) in ethanol (5 L) and heatedat reflux for 15 h. Ethanol was evaporated under reduced pressure, theresidue was diluted with water (1 L) and chilled to 0° C. Concentratedhydrochloric acid (2 L) was slowly added to adjust pH to ˜2, whilemaintaining temperature below 10° C. and stirred for 1 h. Theprecipitated solid was filtered, washed with water (1 L) and pet ether(1 L) and dried under vacuum at 60° C., to afford 550 g (70%) of1,5-Dimethyl-3-phenyl-1H-pyrrole-2,4-dicarboxylic acid D as a whitesolid. [TLC system: ethyl acetate:Pet ether (3:7); R_(f) value: 0.15].

1, 2-Dimethyl-4-phenyl-1H-pyrrole (E)

A suspension of E (550 g, 2.123 mol) in ethanolamine (1.5 L) was heatedto 175° C. (under N₂) and maintained for 1 h. The reaction mixture wascooled to room temperature, diluted with water (500 mL) and extractedwith ethyl acetate (3×200 mL). The combined organic layers were washedsuccessively with water (2×100 mL) and brine (2×100 mL), dried overanhydrous sodium sulfate and concentrated in vacuo below 40° C. to givea crude product. Flash chromatography over neutral alumina using 5%ethyl acetate in pet ether as eluent afforded 280 g (77%) of1,2-dimethyl-4-phenyl-1H-pyrrole E, as a white solid. [TLC system: ethylacetate:Pet ether (3:7); R_(f) value: 0.75].

(1, 5-Dimethyl-3-phenyl-1H-pyrrol-2-yl)-oxo-acetyl chloride (F)

Oxalyl chloride (116 mL, 1.286 mol) was added slowly to a cooledsolution of E (250 g, 1.169 mol) in dry dichloromethane (3×200 mL) at 0°C. The reaction mixture was warmed to room temperature and stirred for 1h. The solvent was evaporated to dryness in vacuo to afford 340 g (89%)of 1,5-dimethyl-3-phenyl-1H-pyrrol-2-yl)-oxo-acetyl chloride F as abrown oily liquid. [TLC system: ethyl acetate:Pet ether (3:7); R_(f)value: 0.65]

4-Nitro phenyl piperazine (G)

A solution of 1-chloro-4-nitro benzene (650 g, 4.140 mol) in diglyme (1L) was added to a solution of piperazine (2.84 Kg, 33.12 mol) in diglyme(500 mL) at 100° C. and the resultant mass was stirred at 100° C. for 6h. The mixture was cooled to 40-45° C., water (5 L) was added; warmed toroom temperature and stirred for 1 h. The precipitated solid wasfiltered, washed with water (1 L), pet ether (500 mL) and dried to give700 g (81%) of 4-nitro phenyl piperazine G as yellow colour solid. [TLCsystem: Ethyl acetate:pet ether (3:7); R_(f) value: 0.70].

5-Fluoro-2-[4-(4-nitro-phenyl)-piperazin-1-yl]-pyrimidine (H)

2-Chloro-5-fluoropyrimidine (281 g, 2.12 mol) was added to suspension of4-nitro phenyl piperazine G (400 g, 1.93 mol) and potassium carbonate(532 g, 3.85 mol) in diglyme (2.5 L), the resulting mixture was stirredat 100° C. for 6 h. On completion the mixture was cooled to 0° C. andfiltered, the solid was taken in water (5 L) and stirred for 30 mins.The suspension was filtered, the solid cake was washed with water (1 L),pet ether (1 L) and dried under vacuum to afford 500 g (85%) of5-fluoro-2-[4-(4-nitro-phenyl)-piperazin-1-yl]-pyrimidine H as yellowcolour solid. [TLC system: Ethyl acetate:pet ether (3:7); R_(f) value:0.70].

4-[4-(5-Fluoro-pyrimidin-2-yl)-piperazin-1-yl]-phenyl amine (I)

A solution of sodium dithionite (1.27 Kg, 7.32 mol) in water (6 L) wasadded to a suspension of H (500 g, 1.83 mol) and sodium bicarbonate (614g, 7.32 mol) in methanol (6 L) at 65° C. The resultant mixture wasstirred at 65° C. for 2 h. The reaction mass was cooled to 10-15° C. andfiltered. The residue was partitioned between water (2 L) and ethylacetate (5 L), the organic layer was washed with water (2 L), brine (2L) and dried over anhydrous sodium sulfate. Concentrated in vacuo toafford 290 g (64%) of4-[4-(5-fluoro-pyrimidin-2-yl)-piperazin-1-yl]-phenyl amine I as solid.[TLC system: Methanol:Chloroform (1:9); R_(f) value: 0.50].

2-(1,5-Dimethyl-3-phenyl-1H-pyrro-2-yl)-N-{4-[4-(5-fluoro-pyrimidin-2-yl)-piperazin-1-yl]-phenyl}-2-oxo-acetamide

A solution of F (332 g, 1.27 mol) in dichloromethane (3 L) was added toa stirred solution of I (290 g, 1.06 mol) and triethylamine (294 mL,2.12 mol) in dichloromethane (3 L) at 0° C. The reaction mixture waswarmed to room temperature and stirred for 30 min. The reaction mixturewas quenched with water and extracted with dichloromethane (6×500 mL).The combined organic layers were washed successively with saturatedsodium bicarbonate solution (1.5 L), water (1 L), brine (1.5 L) andfinally dried over anhydrous sodium sulfate. The organic layer wasstirred with neutral alumina (1 Kg) at room temperature for 30 min andfiltered. The filtrate was concentrated in vacuo to give the crudecompound which on washing with diethyl ether (300 mL) and followed bytrituration with ethanol (3 L) at 80° C. for 1 h and cooled to roomtemperature, filtered, washed with ethanol (500 mL) followed by hexane(200 mL) and dried to give 340 g (64%) of2-(1,5-dimethyl-3-phenyl-1H-pyrro-2-yl)-N-{4-[4-(5-fluoro-pyrimidin-2-yl-piperazin-1yl]-phenyl}-2-oxo-acetamideas yellow color solid. [TLC System: Ethyl acetate:Pet ether (1:1); R_(f)value: 0.65].NMR data for 2-(1,5-Dimethyl-3-phenyl-1H-pyrro-2-yl)-N-{4-[4-(5-fluoro-pyrimidin-2-yl)-piperazin-1-yl]-phenyl}-2-oxo-acetamide(¹H NMR (400 MHz, CDCl₃)) are provided in FIG. 1. The signal wasdetected in the MS spectrum at 499.1 [M+H]⁺.

Example 2: Anti-Fungal Activity of the Compound of Formula I

Data demonstrating that2-(1,5-dimethyl-3-phenyl-1H-pyrrol-2-yl)-N-(4-(4-(5-fluoropyrimidin-2-yl)piperazin-1-yl)phenyl)-2-oxoacetamideinhibits the growth of a wide variety of fungi are presented in theinternational patent application having application numberPCT/GB2015/053546. Information relating to the biological activity ofthe compound of Formula I is incorporated by reference.

PCT/GB2015/053546 describes experiments comparing the antifungalactivity of the compound of Formula I to various reference compounds.The experiments described in PCT/GB2015/053546 show that the compound ofFormula I inhibits growth of fungal organisms with MIC (minimuminhibitory concentrations; i.e. the lowest drug concentration thatinhibits growth of an organism by >80% compared with a drug freecontrol) as follows:

Band MIC/mg/L A MIC ≤ 0.005 B 0.005 < MIC ≤ 0.01 C 0.01 < MIC ≤ 0.02 D0.02 < MIC ≤ 0.04 E 0.04 < MIC ≤ 0.06 F MIC ≥ 0.06 MIC band Organism(RPMI medium) A. niger C A. fumigatus C A. terreus A A. flavus B A.terreus 49 A A. fumigatus 210 C S. apiospermum 13486 D S. apiospermum15848 D S. apiospermum 451 B S. apiospermum 4883 C S. apiospermum 7935 CS. apiospermum 8353 B S. prolificans 18389 C S. prolificans 206 D S.prolificans 6322 D S. species 15849 E S. apiospermum 13486 D S.prolifican 201 D S. prolifican 13486 D S. prolifican 7935 C S.prolifican 15848 D S. prolifican 8353 B S. prolifican 451 B S.prolifican 4883 C S. prolifican 15849 E S. prolifican 1121 A S.apiospermum 1124 A

The compound of Formula 1 showed also good antifungal activity againstS. dehoogii, S. boydii and S. aurantiacum.

PCT/GB2015/053546 also describes experiments in which the compound ofFormula I was tested in vivo in a mouse model. The data in thatapplication shows that the compound of Formula I gives excellentefficacy in murine models of invasive aspergillosis, and that thecompound of Formula I is able to reduce galactomannan indices in A.fumigatus infected mice. The compound of Formula I is also able toincrease survival of mice infected with Lomentospora prolificans FMR3569 relative to control experiments using the anti fungal agentvoriconazole

In the in vivo experiments described in PCT/GB2015/053546, the compoundof Formula I was administered orally by gavage. The administration ofthe compound of Formula I in PCT/GB2015/053546 was not achieved usingthe formulations of the present invention.

Example 3: PK Experiments—In Vivo Rodent Experiments to DeterminePreferred Oral Formulations of the Compound of Formula I

Various formulations were investigated in in vivo PK studies in rats todetermine the optimal oral formulation for dosing to animals and humans.

The compound of Formula I was administered to rats by IV at a dosage of10 mg/kg. The vehicle comprised 15% hydroxypropyl beta cyclodextrin(Kelptose HPB parenteral grade), 5% DMSO and water for injection. Theformulation was filtered using a ≤0.22 μm polyethersulfone (PES)membrane filter prior to use. The dose volume administered was 5 mL/kg.0.2 mL blood samples from the sublingal vein under isofluraneanaesthesia were obtained 5 minutes after IV administration and storedin EDTA. Plasma was separated by centrifugation (1500×g, 10 mins, ca. 4°C.) and was frozen at −70° C. prior to analysis. Analysis to calculateplasma concentration of the compound of Formula I was conducted byLC-MS/MS. Typical AUC values (AUC_(IV)) of 13500 ng·hr/ml were recorded,corresponding to 100% bioavailability (by definition). The AUC_(IV)value was subsequently used to calculate the bioavailability of exampleoral formulations as described above.

Formulation 1

A crystalline sample of the compound of Formula I was formulated inPEG300. The formulation was dosed p.o. (gavage) to rats at 10 mg/kg(dose volume=5 mL/kg). 0.2 mL blood samples from the sublingal veinunder isoflurane anaesthesia were obtained 15 minutes after oraladministration and stored in EDTA. Samples were then taken 0.5, 1, 2, 4,8, 12 and 24 hours post-dose. PK data (determined as for the IVformulation) revealed AUC_(PO) (0-24 hours) of 6610 ng·hr/ml,corresponding to a bioavailability (F) of 49%.

Formulation 2

A crystalline sample of the compound of Formula I was formulated in asolution of 90% PEG300:10% TPGS (d-α-Tocopheryl polyethylene glycol 1000succinate). TPGS is a known bioavailability enhancer that acts as a drugsolubiliser and a precipitation inhibitor and was incorporated as itspresence has proved beneficial in formulations of other insolublecompounds. When dosed p.o. as a solution in a vehicle consisting ofPEG300/TPGS at a dose of 150 mg/kg (formulation concentration; 15mg/mL), the AUC_(PO) (0-24 hours) values in male and female fasted ratswere 34271 ng·hr/ml and 76963 ng·hr/ml respectively, equivalent tobioavailability values of 17% and 38% respectively. However, when thisformulation was administered for extended periods, side effects ofdiarrhea were observed indicating that the formulation would beunsuitable for clinical administration.

Formulations 3 and 4

A crystalline sample of the compound of Formula I was micronised by jetmilling to a final particle size of D(v0.9)=6.7 μm, and subsequentlysuspended in a mixture of HPMC (75%) and SDS (sodium dodecyl sulphate0.05%). This formulation (formulation 3) was dosed p.o. to male andfemale rats at 150 mg/kg. PK experiments gave AUC_(PO) (0-24 hours)values of 9004 ng·hr/ml and 26286 ng·hr/ml for male and female ratsrespectively, equivalent to a bioavailability of 4.5% and 13%respectively.

The compound of Formula I was nanonised in a similar manner andformulated in HPMC/SDS as for formulation 3 (to give formulation 4), andwas administered to male and female rats p.o. PK experiments gaveAUC_(PO) (0-24 hours) values of 17785 ng·hr/ml and 40272 ng·hr/ml inmale and female rats respectively, equivalent to a bioavailability of 9%and 20%, respectively.

Neither formulation 3 nor formulation 4 yielded adequate bioavailabilityof the compound of Formula I. Furthermore, PK experiments revealed thatthe time for maximum plasma concentration of the compound of formula I(T_(max)) was significantly delayed suggesting delayed/incompleteabsorption.

Formulation 5

The compound of Formula I was formulated with hydroxypropyl methylcellulose acetate succinate (HPMCAS) (10% w/w compound of Formula I:90%HPMCAS). The formulation was spray-dried from a solution of 3:1 v/vdichloromethane:methanol. The formulation was administered p.o. to maleand female rats at a dose of 10 mg/kg in 20 mM phosphate buffer(suspension). PK experiments gave AUC_(PO) (0-24 hours) values of 13202ng·hr/ml and 27774 ng·hr/ml in male and female rats, respectively,indicating a bioavailability >95% in both sexes.

Unlike formulation 2, formulation 5 was tolerated by both male andfemale rats with no GI tract disturbances. This allowed toxicologicalstudies to be performed over 1-3 months. Formulation 5 was welltolerated by both male and female rats over this period.

Comparison of Formulation 2 and Formulation 5

A comparison of the results of PK experiments conducted usingformulations 2 and was conducted at equivalent dosages (50 mg/kg) dosedp.o. twice daily (BID) for 28 days. These data show that better exposurewas obtained using formulation 5 as compared to formulation 2, with thefurther benefit of the absence of negative effects on the GI tractassociated with formulation 2.

Cmpd of AUC_(PO) ng · hr/ml Formulation Formula I/Dose Sex Day 1 Day 282 50 mg/kg Male 19880 23280 Female 39400 92390 5 50 mg/kg Male 2886044880 Female 43440 149700

Example 4: PK Experiments—In Vivo Cynomolgus Monkey Experiments toDetermine Preferred Oral Formulations of the Compound of Formula I

Various formulations were investigated in in vivo PK studies incynomolgus monkeys to determine the optimal oral formulation for dosingto animals and humans.

The compound of Formula I was administered to male and female cynomolgusmonkeys by IV at a dosage of 10 mg/kg. The IV formulation was asdescribed in Example 3. PK experiments gave typical AUC values(AUC_(IV)) of 18000 ng·hr/ml. The AUC_(IV) value was subsequently usedto calculate the bioavailability of example oral formulations asdescribed above.

Formulation 2

Studies were conducted using Formulation 2 as described above. Thecompound of Formula I was administered p.o. to male and femalecynomolgus monkeys at a dose level of 10 mg/kg. (dose volume 5 mL/kg). 2mL blood samples were obtained 5 minutes via femoral puncture(vein/artery) into commercially available K₂EDTA tubes using 23 Gneedles coupled to a suitable syringe. Samples were taken 0.25, 0.5, 1,2, 4, 8, 12 and 24 hours after administration of the formulation. Bloodsamples were kept on ice until centrifugation at 1600×g (10 mins, ca. 5°C.) to separate plasma. Analysis to calculate plasma concentration ofthe compound of Formula I was conducted by LC-MS/MS. PK experiments gaveAUC_(PO) (0-24 hours) values of 14915 ng·hr/ml and 8192 ng·hr/ml in maleand female subjects, respectively, indicating a bioavailability of 83%and 46% in males and females, respectively.

Unfortunately, gastrointestinal problems (notably diarrhea) as observedin experiments in rats (see Example 3 above) were noted also forcynomolgus monkeys. Studies of 1 month duration with the PEG/TPGSvehicle showed that diarrhea was a major problem despite the use ofanti-diarrheal medication (Diosmectite; “Smecta”, 1 bag (3 g) per animal4 hours post-dose). These studies showed that despite promising oralbioavailability of the compound of Formula I, formulation 2 was unlikelyto be suitable for clinical administration.

Formulation 5

Studies were conducted using Formulation 5 as described in Example 3.The compound of Formula I was administered p.o. to male and femalecynomolgus monkeys at a dose level of 10 mg/kg (dose volume: 5 mg/kg;formulation concentration 2 mg/mL of the compound of Formula I,corresponding to 20 mg/mL of the spray-dried particles)

The particles produced as described for Formulation 5 were suspended indifferent buffer compositions/volumes. PK experiments as described abovegave AUC_(PO) (0-24 hours) values as follows:

AUC_(PO) (0-24 hours)/ Sex Dose/mg/kg Vehicle ng · hr/ml Male 10 10ml/20 mM phosphate buffer 16804 Female 10 10 ml/20 mM phosphate buffer20871 Male 10  5 ml/40 mM phosphate buffer 26112 Female 10  5 ml/40 mMphosphate buffer 12684

As can be seen, when low buffer salt concentrations (20 mM) and highbuffer volumes (10 mL) were used, or when low buffer volumes (5 mL) buthigher buffer strengths (40 mM) were used, high AUC_(PO) (0-24 hours)values were observed. Under these conditions, bioavailability >90% wasobserved using 10 ml of 20 mM phosphate buffer and bioavailability >70%using 5 ml of 40 mM phosphate buffer. (Slight variation in the resultsobtained using female subjects dosed with 5 ml of 40 mm buffer wasobserved which accounts for the apparently poorer bioavailability inthis subject group).

Studies carried out with formulation 5 at a variety of doses incynomolgus monkeys revealed a complete lack of gastrointestinal sideeffects; the formulation is thus much better tolerated in mammals thanformulation 2 described above. High drug levels were obtained afterrepeat dosing, with improved dose proportionality observed betweendoses.

Comparison of Formulation 2 and Formulation 5

A comparison of the results of PK experiments conducted usingformulations 2 and was conducted at equivalent dosages (50 mg/kg) dosedp.o. twice daily (BID) for 14 days. These data show that better exposurewas obtained using formulation 5 as compared to formulation 2, with thefurther benefit of the absence of negative effects on the GI tractassociated with formulation 2.

Cmpd of AUC_(PO) ng · hr/ml Formulation Formula I/Dose Sex Day 1 Day 142 50 mg/kg Male 50460 59590 Female 32160 91250 5 50 mg/kg Male 5720086745 Female 48810 110500The above data obtained in both rat and cynomolgus monkey shows thatwhen the compound of Formula I is formulated in the form of Formulation5 described above, excellent pharmacokinetics in both rat and cynomolgusmonkey is observed. The formulation is well tolerated and is especiallybeneficial compared with solvent based vehicles such as PEG/TPGS due tothe absence of observed GI symptoms which would be likely to requireintervention with antidiarrheal treatments if used clinically. There isalso better dose proportionality in AUC and Cmax using this formulationcompared with solvent based formulations.

Formulation 6

The compound of Formula I was formulated in a solution of 25% DMSO:75%PEG300 (% v/v) (concentration of compound in vehicle: 2 mg/mL). Theformulation was administered p.o. to male and female cynomolgus monkeysat a dose of 10 mg/kg (administered dose volume: 5 mL/kg). 2 mL bloodsamples were obtained 5 minutes via femoral puncture (vein/artery) intocommercially available K₂EDTA tubes using 23 G needles coupled to asuitable syringe. Samples were taken 0.25, 0.5, 1, 2, 4, 8, 12 and 24hours after administration of the formulation. Blood samples were kepton ice until centrifugation at 1600×g (10 mins, ca. 5° C.) to separateplasma. Analysis to calculate plasma concentration of the compound ofFormula I was conducted by LC/MS-MS. PK experiments gave AUC_(PO) (0-24hours) values of 2900 ng·hr/ml and 1480 ng·hr/ml in male and femalesubjects, respectively, indicating a bioavailability of 16% and 8% inmales and females, respectively.

Formulation 7

A lipid based formulation (Catalent) intended to assist solubilisation(and thus increase bioavailability) of the compound of Formula I wasdeveloped and tested in cynomolgus monkeys as described above. Thisformulation was administered p.o. to male and female cynomolgus monkeysat a dose of 10 mg/kg. PK experiments gave AUC_(PO) (0-24 hours) valuesof 3282 ng·hr/ml and 1924 ng·hr/ml in males and females, respectively,indicating very low bioavailability values of 18% and 11%, respectively.

Example 5: PK Experiments—In Vivo Human Trials

Studies were conducted using a spray-dried formulation in accordancewith the compositions of the inventions. The formulation consisted of20% of the compound of Formula I/80% HPMCAS suspended in 20 mM phosphatebuffer as described for formulation 5 in Example 4. Blood samples weretaken following dosage of the formulation and plasma concentrations ofthe compound of Formula I were determined as described above.

Initial experiments were conducted to determine the effect of varyingthe administered dose of the formulation. Results are presented in thefollowing Table.

Dose T_(max) ^(#) C_(max) AUC₀₋₁₂ AUC₀₋₂₄ AUC_(0-t) AUC_(0-∞) t_(1/2)CL/F C₁₂ ⁺ C₂₄ ⁺ MRT V_(ss)/F (mg/kg) (h) (μg/mL) (μg · h/mL) (h)(mL/h)/kg (μg/mL) (h) (L/kg) 2 1.25 0.913 4.36 6.84 11.68 12.18 23.3 1900.154 0.205 29.5 5.23 4 2.5 2.30 13.30 19.24 40.51 46.67 37.0 112 0.5520.547 47.6 4.45 6 3.5 3.67 19.85 32.09 64.72 69.44 29.2 95 0.814 1.01840.7 3.79 8 4.0 5.32 28.38 41.76 81.86 88.20 32.3 94 1.290 1.254 44.14.05 10 3.0 5.79 28.80 44.14 79.21^($) 99.14 31.1 111 1.352 1.338 42.04.34 ^(#)Median values. ⁺Concentrations at 12 and 24 h post-dose (forpotential repeat dose purposes). ^($)AUC_(0-t) values only from 0-72 hat present to prevent unblinding.In the preceding table: T_(max)=time of observed maximum concentrationof the compound of Formula I; C_(max)=observed maximum concentration;AUC_(A-B)=area under the PK plasma curve vs time curve, wherein A istime=0 hours and B=time to which curve determined; t_(1/2)=apparentterminal elimination half-life; CL=clearance; C_(n) (n=12,24)=concentration of compound of Formula I at indicated time point;MRT=mean residence time; V_(SS)=volume of distribution at steady-state

Plots of C_(max) and AUC_(0-∞) as a function of administered doserevealed good dose proportionality (C_(max): proportionality=1.18; AUC:proportionality=1.30) as shown in FIGS. 1 to 3. Excellentbioavailability of ca. 95%+ was observed.

1. A pharmaceutical composition suitable for oral administration whereinthe composition comprises spray-dried particles of a compound of FormulaI or a pharmaceutically acceptable salt thereof

2-(1,5-dimethyl-3-phenyl-1H-pyrrol-2-yl)-N-(4-(4-(5-fluoropyrimidin-2-yl)piperazin-1-yl)phenyl)-2-oxoacetamide2. A pharmaceutical composition according to claim 1 comprisingspray-dried particles of a compound of Formula I.
 3. A pharmaceuticalcomposition according to claim 1 or claim 2 wherein the compound ofFormula I is substantially amorphous.
 4. A pharmaceutical compositionaccording to any one of the preceding claims which further comprises oneor more excipients.
 5. A pharmaceutical composition according to claim4, wherein the composition comprises the excipient hydroxypropyl methylcellulose acetate succinate (HPMCAS).
 6. A pharmaceutical compositionaccording to any one of claim 4 or 5 wherein the mass ratio of thecompound of Formula I to excipient is from 1:100 to 1:1.
 7. Apharmaceutical composition according to any one of claims 4 to 6 whereinthe mass ratio of the compound of Formula I to excipient is from 1:15 to1:2.
 8. A pharmaceutical composition according to any one of theprevious claims wherein the particles are obtainable by spray-dryingfrom a solution comprising an organic solvent selected fromdichloromethane, methanol, and mixtures thereof.
 9. A pharmaceuticalcomposition according to any one of the previous claims and furthercomprising one or more pharmaceutically acceptable binders and/orcarriers and/or excipients and/or diluents and/or adjuvants.
 10. Apharmaceutical composition according to any one of the previous claimsin the form of a solid oral dosage form.
 11. A pharmaceuticalcomposition according to any one of the previous claims in the form of aliquid oral dosage form.
 12. A pharmaceutical composition according toclaim 11 wherein the liquid oral dosage form further comprises apharmaceutically acceptable buffer having a pKa in the range 6.0 to 8.0.13. A pharmaceutical composition according to claim 11 or claim 12wherein the buffer is from 1 mM to 200 mM phosphate buffer and whereinthe composition is buffered to about pH
 7. 14. A pharmaceuticalcomposition suitable for parenteral administration comprising (i) acompound of Formula I or a pharmaceutically acceptable salt thereof,

2-(1,5-dimethyl-3-phenyl-1H-pyrrol-2-yl)-N-(4-(4-(5-fluoropyrimidin-2-yl)piperazin-1-yl)phenyl)-2-oxoacetamide(ii) cyclodextrin or modified cyclodextrin, and (iii) polyethyleneglycol.
 15. A pharmaceutical composition according to claim 14comprising: from 10 wt % to 40 wt % of cyclodextrin or modifiedcyclodextrin; and/or from 10 wt % to 40 wt % of polyethylene glycol. 16.A pharmaceutical composition according to claim 14 or claim 15 whereinthe cyclodextrin or modified cyclodextrin is hydroxy propyl betacyclodextrin.
 17. A pharmaceutical composition according to any ofclaims 14 to 16 wherein the polyethylene glycol is PEG300 or PEG400. 18.A pharmaceutical composition according to any one of claims 14 to 17which further comprises polyvinyl pyrrolidone (Povidone).
 19. Apharmaceutical composition according to any one of claims 14 to 18wherein the compound of Formula I or the pharmaceutically acceptablesalt thereof is present at a concentration of from 1 mg/mL to 10 mg/mL.20. A pharmaceutical composition according to any one of claims 14 to 19which further comprises one or more pharmaceutically acceptable carriersand/or excipients and/or diluents and/or adjuvants.
 21. A pharmaceuticalcomposition according to any one of claims 14 to 20 comprising: 4 mg/mL(relative to the final volume of the composition) of the compound ofFormula I 25 wt % of hydroxy propyl beta cyclodextrin; wt % of PEG400; 1wt % of polyvinyl pyrrolidone (Povidone); phosphoric acid in sufficientamount to adjust the pH of the pharmaceutical composition to pH 5.0; andwater to 100%.
 22. A pharmaceutical composition according to any one ofthe previous claims for use in a method of treatment of a human oranimal subject in need thereof.
 23. A composition for use according toclaim 22 wherein the treatment comprises prevention of or treatment offungal infection in the subject.
 24. A method of preventing or treatingfungal infection in a human or animal subject in need thereof, saidmethod comprising administering to the human or animal subject atherapeutically effective amount of a pharmaceutical compositionaccording to any one of claims 1 to
 21. 25. Use of a pharmaceuticalcomposition according to any one of claims 1 to 21 in the manufacture ofa medicament for use in the prevention or treatment of fungal infectionin a human or animal subject in need thereof.
 26. A method of producinga pharmaceutical composition comprising a compound of Formula I or apharmaceutically acceptable salt thereof

2-(1,5-dimethyl-3-phenyl-1H-pyrrol-2-yl)-N-(4-(4-(5-fluoropyrimidin-2-yl)piperazin-1-yl)phenyl)-2-oxoacetamidewherein said method comprises spray-drying a solution of the compound ofFormula (I) or salt thereof.
 27. A method according to claim 26comprising: i) dissolving one or more excipients in a solvent; ii)adding the compound of Formula I to the solution produced in step (i);and iii) spray drying the solution produced in step (ii).
 28. A methodaccording to claim 27 wherein: the excipient is hydroxypropyl methylcellulose acetate succinate (HPMCAS); the solvent is a mixture ofdichloromethane and methanol, wherein the volume ratio ofdichloromethane to methanol is from 5:1 to 1:1; the concentration of theexcipient in the solvent is from 5% to 20% w/v; and the compound ofFormula I is added to the solution of the excipient in the solvent togive a concentration of 0.5% to 10% by mass.
 29. A pharmaceuticalcomposition suitable for oral administration wherein the compositioncomprises substantially amorphous particles of a compound of Formula Ior a pharmaceutically acceptable salt thereof.